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Abstract
Metallic nanoparticles (NPs) especially silver (Ag) NPs have shown immense potential in medical applications due to their distinctive physio-chemical and biological properties. This article reports the conjugation of Ag NPs with Rubus fairholmianus extract. The modification of Ag NPs was confirmed using various physico-chemical characterization techniques. The cytotoxic effect of Rubus-conjugated Ag NPs (RAgNPs) was studied by LDH assay and proliferation by ATP assay. The apoptotic inducing ability of the NPs were investigated by Annexin V/PI staining, caspase 3/7 analysis, cytochrome c release, intracellular ROS analysis, Hoechst staining and mitochondrial membrane potential analysis using flow cytometry. The expression of apoptotic proteins caspase 3, Bax and P53 were analyzed using ELISA and caspase 3, Bax using western blotting. Cells treated with 10 µg/mL RAgNPs showed an increased number of cell death by microscopic analysis compared to untreated control cells. The RAgNPs induced a statistically significant dose-dependent decrease in proliferation (p < 0.001 for 5 and 10 µg/mL) and increased cytotoxicity in MCF-7 cells. A 1.83 fold increase in cytotoxicity was observed in cells treated with 10 µg/mL (p < 0.05) compared to the untreated cells. Nuclear damage and intracellular ROS production were observed upon treatment with all tested concentrations of RAgNPs and the highest concentrations (5 and 10 µg/mL) showed significant (p < 0.05, p < 0.01) expression of caspase 3, Bax and P53 proteins. The data strongly suggest that RAgNPs induces cell death in MCF-7 cells through the mitochondrial-mediated intrinsic apoptosis pathway. The present investigation supports the potential of RAgNPs in anticancer drug development.
Highlights
Cancer is one of the most deadly diseases with high mortality rates
The amorphous region from 12° to 30° belongs to the Rubus fairholmianus root acetone extracts (RFRA) extracts as it contains various organic moieties and indicates the crystallisation of bioorganic phase exist on the surface of Ag NPs
We have examined the expression of caspase 3 and subsequent apoptosis in MCF-7 cells treated with Rubus-conjugated Ag NPs (RAgNPs)
Summary
Cancer is one of the most deadly diseases with high mortality rates. It refers to a collection of diseases where certain cells the body begin to grow and multiply disorderly. The chemi-synthetic process leads to the presence of toxic chemicals on the surfaces, which have harmful effects in medical applications[15] This problem can be overcome by green synthesis of NPs16. There is a lack of detailed research results regarding the anticancer effects of green synthesized Ag NPs. It will be of interest to determine the synergistic cytotoxic effect of Ag NPs and Rubus extract on cancer cells as they could demonstrate better biocompatibility and enhanced therapeutic attributes. To the authors knowledge no study has been undertaken to determine the in vitro anti-proliferative effects of Ag NPs capped with Rubus extracts against MCF-7 breast carcinoma cells. The current study was performed to modify Ag NPs with Rubus extract, and evaluate its anticancer potential against MCF-7 cells in vitro. The cytotoxicity, proliferation and caspase mediated apoptotic effects of capped Ag NPs were monitored through various in vitro analyses
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